Objective To understand the current situation as well as the implementation of infection prevention and control measures against bacterial contamination of external and vaginal ultrasound probes in medical institutions (MIs), and explore the risk of ultrasound probe-related infection under current prevention and control measures. Methods Through on-site sampling, external ultrasound probes from 179 MIs and vaginal ultrasound probes from 83 MIs in 15 prefecture-level cities in Henan Province underwent quantitative bacterial detection, and the implementation of relevant infection prevention and control measures was assessed using questionnaire survey. Results A total of 513 ultrasound probe specimens were detected. Bacterial detection rate and exceedance rate of the disinfected vaginal ultrasound probes were 18.02% (20/111) and 2.70% (3/111), respectively, which were both lower than external ultrasound probes (40.05%[161/402] and 5.22%[21/402], respectively). Bacterial detection rate of disinfected specimens was lower than that of the towel-wiping group ([33.86%, 149/440] vs[50.00%, 27/54], χ²=5.46, P=0.019). Bacterial detection rate of the quaternary ammonium salt disinfectant group was lower than that of the alcohol disinfectant group ([17.31%, 18/104] vs[40.63%, 128/315], χ²=18.74, P<0.001). The qualified rates of specimens using different disinfection methods and with different probe types showed no statistically significant differences (all P>0.05). The implementation rates of "single disinfection for every use" for external ultrasound probes and vaginal ultrasound probes were 60.34% (108/179) and 70.54% (79/112), respectively. The disinfectants used were mainly quaternary ammonium salts (57.27%) and alcohols (36.36%). The coupling agents used for external and vaginal ultrasound examinations were mainly large packaged products for multiple time use, accounting for 93.85% and 57.89%, respectively. 33.33% (56/168) of external ultrasound coupling agents and 20.00% (11/55) of vaginal ultrasound coupling agents were used for more than 7 days. In vaginal ultrasound exa-minations, the usage rates of disinfectant coupling agents and sterile coupling agents were 28.42% (27/95) and 14.74% (14/95), respectively. In vaginal ultrasound examinations, 50.00% (56/112) of MIs used condoms as probe co-vers, and 45.54% (51/112) used sound-transmission ultrasound isolation membranes. 60.55% (66/109) of MIs didn’t change gloves after cleaning and disinfecting the vaginal ultrasound probe and before examination. Conclusion Ultrasound probes have cross infection risks in key processes such as reprocessing (disinfection/sterilization), phy-sical barrier use, coupling agent selection, and glove replacement. It is urgent to develop and implement standar-dized ultrasound probe infection prevention and control technical specifications based on evidence-based research.